Search Results (370)

Orchid root-associated fungal endophytes (ORAFEs) represent an important component of fungal biodiversity with potential applications in orchid conservation and sustainable production. ORAFEs were isolated from the roots of four orchid species in Veracruz, Mexico: Gongora galeata, Maxillaria densa, Maxillaria tenuifolia, and Maxillaria variabilis to investigate their diversity and plant-growth-promoting (PGP) properties. Taxonomic identification relied on morphological and molecular phylogenetic analyses using ITS, TEF1-α, β-tubulin (TUB2), and actin (ACT). Fifty-six isolates were grouped into 20 morphotypes representing 15 genera, including Aspergillus, Biscogniauxia, Chaetomium, Colletotrichum, Thelonectria, Diaporthe, Fusarium, Geotrichum, Hypoxylon, Lasiodiplodia, Mucor, Periconia, Pestalotiopsis, Trichoderma, Xylaria, and two isolates that may represent undescribed taxa requiring further polyphasic taxonomic investigation. Functional experiments indicated a wide range of PGP activities: ten strains solubilized phosphate, eight produced siderophores, twelve synthesized indole-3-acetic acid, fourteen produced pectinases, six fungi exhibited antibacterial activity against Bacillus subtilis, and five inhibited Staphylococcus aureus. Furthermore, Trichoderma atroviride TR1-T2 and Xylaria multiplex XY1-D42 showed antagonistic activity against Phytopythium vexans, characterized by a contact-dependent displacement effect. Five strains were chosen for further research as orchid bioinoculants based on their PGP and antagonistic traits: Chaetomium cochliodes CH1-V10, Mucor pseudolusitanicus MU2-T6, Thelonectria veuillotiana TH1-T3, Trichoderma atroviride TR1-T2, and Xylaria multiplex XY1-D42. These endophytes have the potential to improve in vitro propagation and facilitate ex vitro acclimatization of orchids, highlighting their importance in sustainable orchid production. Full article

Adenium obesum (Forssk.) Roem. & Schult (desert rose) is one of the most popular ornamental species cultivated in residential gardens and as a potted plant for balconies and verandas in Brazil. However, Neocosmospora keratoplastica has recently been identified as one of the causal agents of rot disease in this species. Therefore, this study evaluated the antifungal potential of Ocimum tenuiflorum L. essential oil (EO) as a preventive and curative treatment against this pathogen, while assessing its phytotoxicity and selectivity toward beneficial fungi. The EO was obtained by hydrodistillation and chemically characterized. In vitro mycelial growth inhibition assays, in vivo evaluations on A. obesum seedlings, and selectivity tests against Trichoderma asperellum were performed. In addition, in silico molecular docking analyses were conducted to investigate the interaction of the major EO constituents with fungal target proteins. Eugenol (47.04%) and 1,8-cineole (29.47%) were identified as the predominant compounds in the EO. Mycelial growth inhibition reached a maximum of 59% at 100 μL mL⁻¹, but strong phytotoxicity was evident; while a concentration of 50 μL mL⁻¹ yielded an 18% inhibition rate representing an approximately three-fold greater effect than that of thiophanate methyl (7.68%) at the recommended dose, with a slight phytotoxicity. The lowest Area Under the Disease Progress Curve (AUDPC) was recorded when the EO was applied preventively, and intermediate concentrations were selective toward T. asperellum. Molecular docking analyses indicated that eugenol and 1,8-cineole exhibited greater affinity for CYP51A than for α/β-tubulin, whereas dehydroaromadendrane and α-copaene showed the strongest interactions with these target proteins. Overall, O. tenuiflorum EO exhibits promising potential as a preventive treatment against N. keratoplastica in desert rose, while preserving beneficial fungi at moderate concentrations. Full article

Background/Objectives: Tubulins (Tub) are central components of microtubules, but intraspecific variation and developmental expression of the Tub family in Gossypium barbadense remain poorly characterized. This study aimed to characterize the GbTub family using a pan-genome framework and identify candidates associated with fiber development and strength. Methods: A total of 50 GbTub genes were identified in the G. barbadense 3-79 reference genome, and their orthologous presence–absence patterns were subsequently assessed across 12 additional G. barbadense accessions. Phylogenetic, presence–absence variation (PAV), Ka/Ks, structural variation (SV), RNA-seq, RT-qPCR, co-expression, and GO enrichment analyses were integrated. Results: Among the 50 reference-defined GbTub genes, 43 were classified as core genes, 6 as near-core genes, and 1 as an accessory gene, and the encoded proteins were classified into α-, β-, and γ-tubulin clades. All genes showed Ka/Ks < 1. Twenty-three GbTub genes differed between the fiber-strength-contrasting accessions 5917 and PimaS-7, and representative expression trends were supported by RT-qPCR. Network analysis prioritized 10 GbTub candidates based on degree centrality. GbTub21 was the sole SV-associated GbTub gene displaying significant differential expression between accessions harboring versus lacking the corresponding SV. Non-Tub neighbors of the candidate hub genes were enriched for cytoskeletal, intracellular-transport, and plasma-membrane functions. Conclusions: The pan-genome analysis reveals strong conservation with limited intraspecific variation in the GbTub family. Co-expression profiles nominate candidates associated with fiber secondary-wall development, and their causal contribution to fiber strength awaits functional dissection. Full article

Background/Objectives: Docetaxel is a frontline chemotherapeutic agent for breast cancer; however, therapeutic resistance remains a major clinical challenge. Emerging evidence suggests that chemotherapy-induced adaptive phosphorylation events can promote survival signaling and contribute to drug resistance. However, the phosphoproteomic mechanisms underlying docetaxel-induced adaptive responses remain poorly characterized. Methods: We performed integrated quantitative proteomic and phosphoproteomic profiling in breast cancer cells following docetaxel exposure. Candidate kinases associated with phosphorylation remodeling were identified and validated using TCGA-BRCA and CPTAC clinical datasets. The functional significance of PAK4 phosphorylation was validated using phosphomimetic and phospho-deficient mutants, pharmacological inhibition, and assessment of microtubule stabilization. Results: Integrated phosphoproteomic analysis revealed extensive phosphorylation remodeling following docetaxel treatment and identified PAK4 as a candidate kinase associated with the adaptive response. Analysis of the CPTAC phosphoproteomic dataset showed that phosphorylation of PAK4 at S474 was elevated in breast cancer tissues, increased with tumor stage, and was associated with poorer overall survival. In breast cancer cells, docetaxel induced phosphorylation of PAK4 at S474 without altering total PAK4 expression. Functionally, phosphomimetic PAK4 (S474D) reduced docetaxel sensitivity, whereas phospho-deficient PAK4 (S474A) enhanced drug sensitivity. Pharmacological inhibition of PAK4 using LCH-7749944 significantly enhanced the inhibitory effect of docetaxel on cell viability and increased apoptosis in breast cancer cells. Mechanistically, PAK4 inhibition enhanced docetaxel-induced microtubule stabilization, as evidenced by increased α-tubulin acetylation and accumulation of stabilized microtubule structures. Conclusions: Our study demonstrates that docetaxel induces global phosphorylation network reprogramming in breast cancer cells and identifies PAK4 S474 phosphorylation as a key determinant of docetaxel sensitivity. Inhibition of PAK4 enhances microtubule stabilization and improves the efficacy of docetaxel, providing a potential combinatorial strategy to overcome taxane resistance. Full article

Reliable reference genes are critical for ensuring the accuracy of RT-qPCR-based gene expression analysis, especially under environmental stress conditions. In this study, Leymus chinensis was used as the experimental material, and eight candidate reference genes—alpha-tubulin (TUA), beta-tubulin (TUB), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), elongation factor 1-alpha (EF1α), 18S ribosomal RNA (18S rRNA), adenylyl cyclase-associated protein (CAP), adenine phosphoribosyl transferase (APRT), and actin (ACT)—were selected to evaluate their expression stability under cold, drought, heat, NaCl, high pH, wounding, abscisic acid (ABA) and jasmonic acid (JA) treatments. Primer specificity and amplification efficiency were first assessed, and the candidate genes were then comprehensively analyzed using geNorm, NormFinder, BestKeeper, and RefFinder. The results showed that the amplification efficiencies of all primers ranged from 95.0% to 107.2%, and the Ct values of the candidate genes ranged from 17.31 to 30.99. Comprehensive analysis using RefFinder showed that ACTIN was the most stable gene under ABA and NaCl treatments, EF1α under heat and wounding treatments, CAP under JA and high pH treatments, APRT under cold treatment, and TUB under drought treatment. geNorm analysis indicated that two reference genes were sufficient for accurate normalization under each treatment condition. The reliability of the screening results was further confirmed by expression-level validation of LcbZIP46, LcWRKY5, and LcFIN1. This study provides a stable reference gene system for RT-qPCR-based expression analysis in Leymus chinensis under environmental stress conditions. Full article

Background: Two novel copper(II) coordination complexes, PH-Cu [(1,10-phenanthroline)(malonato)copper(II)] and PC-Cu [(1,10-phenanthroline)(cyclobutane-1,1-dicarboxylato)copper(II)], were synthesized and evaluated as potential anticancer agents, aiming to characterize structural properties, explore antiproliferative activity and generate mechanistic hypotheses through experimental and computational approaches. Methods: Complexes were characterized by EPR, FTIR-ATR, and ESI-MS, with preliminary SC-XRD data for PH-Cu. Antiproliferative activity was evaluated against six human cancer cell lines using the MTT assay (24 h). Subcellular effects were assessed by fluorescence microscopy and RT-qPCR. Computational studies included DFT geometry optimization, target prediction, molecular docking, and ADMET profiling. Results: Based on spectroscopic and spectrometric data and comparison with analogous Cu(II) complexes, a distorted square-pyramidal coordination geometry was proposed; this assignment was not confirmed by SC-XRD. Both complexes exhibited potent antiproliferative activity, with PH-Cu showing the highest potency in HeLa cells (IC₅₀ = 4.22 µM). Under the same conditions, cisplatin showed substantially lower activity (HepG2: 191.1 µM; Caco-2: 129.6 µM; NCI-H69: >333.3 µM; HeLa: 21.9 µM). Fluorescence microscopy at 18 h revealed pyknosis, karyorrhexis, and microtubule disorganization, consistent with regulated cell death. RT-qPCR of PH-Cu indicated intrinsic apoptotic pathway engagement (BAX +3.20-fold; BCL2 to 0.39-fold of control). DFT-optimized bond lengths were consistent with crystallographic data for analogous complexes. Molecular docking suggested PRKCG, RELA (p65), Caspase-3, and α/β-tubulin as interaction candidates, while ADMET profiling predicted favorable intestinal absorption (>92.8%) and low BBB permeability. Conclusions: These results suggest that the [Cu(phen)] unit constitutes the primary pharmacophore, with the dicarboxylate co-ligand as a modulator of the antiproliferative profile, suggesting promising anticancer pharmacological potential. Full article

Experimental evidence has shown that gallic acid (GA), a naturally occurring polyphenolic compound, and docetaxel (DTX), a taxane chemotherapeutic agent, each possess antitumor activity against multiple cancer types. Although both compounds have been investigated individually, their combined effects in triple-negative breast cancer (TNBC) have received limited attention, and the molecular basis of their interaction remains unclear. The present study examined the in vitro effects of GA and DTX in MDA-MB-231 TNBC cells while simultaneously assessing their comparative cytotoxicity in HaCaT human keratinocytes. Evaluation of treatment efficacy included measurement of cell viability by the MTT assay and assessment of drug interactions using the Chou–Talalay combination index (CI) method. Apoptosis together with cell-cycle distribution was subsequently examined using both Annexin V/PI flow cytometry and TALI^® image-based cytometry. Additional analyses included β-tubulin immunofluorescence (IF), caspase-9 immunocytochemistry, ELISA, wound-healing assays, quantitative real-time PCR, and bioinformatic analyses to investigate treatment-associated biological alterations. Combined exposure to GA and DTX produced a significant reduction in cell viability and exhibited synergistic activity in MDA-MB-231 cells. The coordinated biological response to the combined treatment was characterized by increased apoptotic cell death, arrest of the cell cycle at the G2/M phase, extensive disorganization of the β-tubulin network, and enhanced caspase-9 immunoreactivity. Beyond its effects on cell survival, the combined regimen substantially decreased the release of IL-6, IL-8, and TNF-α, limited wound-healing capacity, and reshaped the expression profile of the apoptosis- and cell cycle-related genes BCL2, BAX, CASP9, and CDKN1A. Bioinformatic analyses further revealed enrichment of apoptosis- and cell-cycle-associated pathways that were generally consistent with the experimental observations. The overall pattern of experimental responses indicates that combining GA with DTX enhances the in vitro antitumor efficacy of DTX in TNBC cells by simultaneously influencing apoptotic pathways, cell-cycle regulation, inflammatory cytokine secretion, and cellular migratory capacity. Although the bioinformatic findings provide supportive hypothesis-generating evidence, additional studies using three-dimensional models, in vivo experiments, and functional validation approaches are necessary to confirm the underlying molecular mechanisms and to further define the translational potential of this therapeutic combination. Full article

Tubulins are essential cytoskeletal components involved in plant cell division and expansion, yet their evolutionary dynamics across plant lineages and potential roles in horticultural seed/kernel development remain insufficiently understood. Here, we identified 2535 tubulin-related genes from 97 plant genomes and performed an integrated phylogenomic analysis. Phylogenetic and synteny network analyses resolved four ancient clades, including α-, β-,γ-tubulin and FtsZ, all of which were predominantly subjected to purifying selection. The α- and β-tubulin subfamilies exhibited lineage-specific expansion in angiosperms, particularly in eudicots, and these expansions were associated with ancient WGD and WGT events while retaining relatively conserved chromosomal contexts. By employing a pyramid-structured microsynteny framework across 12 Rosaceae genomes, we further traced the orthologous conservation and lineage-specific rearrangements of tubulin loci, with Prunus armeniaca as a reference. Spatiotemporal transcriptome profiling of Siberian apricot and kernel apricot revealed a group of tubulin genes highly expressed during key stages of kernel development, highlighting PaTUA1 as a priority candidate gene. Transient overexpression of PaTUA1 in wounded developing apricot kernels was associated with short-term increases in average phytohormone concentrations, including IAA, GA₃, BR, and cytokinins. Together, these results suggest that PaTUA1 represents a promising candidate gene associated with hormone-related responses during apricot kernel development, providing a basis for future functional validation rather than direct evidence of kernel-size determination. Full article

The longissimus dorsi (LD) and biceps femoris (BF) muscles are important meat-producing regions in camels. Investigating differences in meat quality and proteomic profiles between the LD and BF muscles in Junggar Bactrian camels can provide a molecular basis for regulating camel meat quality and genetic improvement. In this study, 20 healthy adult male Junggar Bactrian camels were selected. Following slaughter, muscle samples were collected from the splenius (SP), triceps brachii (TB), LD, external oblique (EO), gluteus medius (GM), and BF. Meat quality parameters (pH, meat color, shear force, drip loss, and cooking loss) were measured. The LD exhibited the highest meat quality among the six cuts, in contrast to the BF, which showed the lowest. Proteomic analysis of LD and BF from 6 Junggar Bactrian camels was conducted to identify proteins associated with meat quality, yielding 81 differentially expressed proteins (DEPs). Gene Ontology (GO) enrichment analysis highlighted several significantly enriched terms among the DEPs (p < 0.05), including calcium-dependent phospholipid binding, zinc ion binding, and metal ion binding. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis (p < 0.05) further indicated notable enrichment in cytoskeletal organization, 2-oxocarboxylate metabolism, and the citric acid cycle. DEPs associated with meat quality were identified, including tubulin α-chain-like 3 and synaptic function regulator FMR1 isoform X15, which can serve as candidate DEPs for shear force. Protein phosphatase 1 regulatory subunit 14C isoform X1 can serve as a candidate differentially expressed protein for pH. Protein phosphatase 1 regulatory subunit 14C isoform X and anchoring protein repeat domain 1 can serve as candidate DEPs for cooking loss. Membrane-associated protein A4 and membrane-associated protein A7 isoform X1, as well as the transcriptional activator of cytochrome c oxidase 1, can serve as candidate DEPs for color a*. These data may serve as a reference for further studies on how different cuts affect meat quality and for practical efforts to improve camel meat quality. Full article

The voltage-dependent anion channel (VDAC) is the primary conduit for ion and metabolite transport across the mitochondrial outer membrane. Positioned at the interface between the cytosol and the mitochondrial compartment, VDAC is uniquely accessible to proteins on both sides of the membrane, making it an interaction hub whose biophysical properties and signaling functions are shaped by protein complexation in addition to its intrinsic pore specialization. Mammals express three isoforms—VDAC1, VDAC2, and VDAC3—sharing a conserved β-barrel scaffold with about 70% identity. However, minor differences in the sequence lead to drastic changes in VDAC isoform affinity with other proteins. Here, we review the molecular mechanisms and physiological consequences of VDAC complexation with a set of well-characterized partners: hexokinase, dimeric tubulin, α-synuclein, mitochondria-associated membrane proteins, B-cell lymphoma 2 (BCL-2) family proteins, and the translocase of the outer membrane (TOM) protein import complex. For each complex, we evaluate the available structural, biophysical, and genetic evidence for isoform specificity, highlight where mechanistic understanding is most advanced, and identify open questions. A consistent principle emerges across all complexes: functionally nonredundant isoform contributions are primarily governed by differential partner affinity and complexation, rather than by differences in pore architecture alone. This framework has direct implications for mitochondria-associated pathologies, including cancer, cardiovascular disease, and neurodegeneration, as well as for the rational design of VDAC-targeting therapeutics. Full article

Background/Objectives: Chronic rhinosinusitis with nasal polyps (CRSwNP) is associated with epithelial remodeling, impaired mucociliary clearance, and altered nitric oxide (NO) metabolism. However, cell type-specific mechanisms underlying epithelial NO signaling remain poorly understood. This study investigated NO-related signaling in differentiated human sinonasal epithelial cells. Methods: Human sinonasal tissues were obtained from patients with CRSwNP (n = 20) and control subjects (n = 20). Air–liquid interface (ALI) cultures were established from donor-derived epithelial cells. Ciliated and non-ciliated cells were identified by immunostaining for acetylated α-tubulin and BCAM. Expression of inducible nitric oxide synthase (iNOS) and endothelial nitric oxide synthase (eNOS) was analyzed by quantitative RT-PCR. Intracellular NO-related fluorescence signals were evaluated using DAF-FM fluorescence imaging. Results: CRSwNP tissues exhibited significantly increased iNOS expression and elevated iNOS/eNOS ratios, whereas eNOS expression did not differ significantly from that in controls. ALI cultures reproduced differentiated sinonasal epithelium containing both ciliated and non-ciliated cell populations. DAF-FM fluorescence signals were significantly higher in ciliated cells than in non-ciliated cells (80.3 ± 25.3 vs. 49.3 ± 21.1). Non-selective NOS inhibition markedly reduced fluorescence signals in both cell types, whereas selective iNOS inhibition reduced but did not abolish signals in ciliated cells. Conclusions: NO-related signaling appears to differ among epithelial cell subtypes. Persistence of fluorescence signals after selective iNOS inhibition suggests a contribution of constitutive NOS activity in ciliated cells, whereas non-ciliated cells appear to rely more heavily on iNOS-dependent pathways. These findings support the hypothesis that altered epithelial NO signaling contributes to epithelial dysfunction and impaired mucociliary homeostasis in CRSwNP. Full article

Melanoma is a highly aggressive and invasive form of skin cancer that arises from the uncontrolled growth of melanocytes. It is characterized by early spread through the lymphatic system and metastasis. The success of metastasis is linked to the ability of melanoma and other cancer cells to resist anoikis, a type of cell death that occurs when cells lose their adhesion to the extracellular matrix. Redox signaling plays an essential role in anoikis resistance. The balance between intracellular levels of nitric oxide (NO) and the reactive oxygen species (ROS) O₂⁻ and H₂O₂ stimulate signaling pathways related to proliferation and survival or cell death. A375 and SK-MEL-28 human melanomas cell lines, representing primary melanoma and lymph node metastatic melanoma cells, respectively, under suspension and adherent culture conditions were used to investigate the redox regulation of anoikis resistance. Both cell lines express the three isoforms of nitric oxide synthases (NOS) and NADPH oxidase 4 (NOX4) as endogenous sources of NO and ROS, respectively. When A375 cells in suspension were treated with the pan-NOS inhibitor L-NAME, their viability decreased. The treatment resulted in a decrease in FAK phosphorylation at Tyr397 and in ERK 1/2 phosphorylation. The expression of FAK, ERK 1/2, β-actin, and α-tubulin were significantly reduced. Treatment with L-NAME led to an increase in the expression of the metalloprotease MMP-9. SK-MEL-28 cells in suspension and treated with the NOX4 inhibitor, GKT36901, exhibited reduced viability. This was accompanied by the inhibition of FAK phosphorylation at Tyr397, ERK 1/2 phosphorylation, and a reduction in the expression of FAK, ERK 1/2, β-actin, and α-tubulin, with a slight elevation in the expression of MMP-9. Migration and invasion were strongly inhibited in A375 cells upon treatment with L-NAME, while treatment with GKT36901 had a marginal effect on the migration and invasion capacities of SK-MEL-28 cells. In summary, melanoma cells employ nitrosative and oxidative stress to shield themselves from anoikis. Nitric oxide was essential for melanoma cells at the primary site for resisting anoikis, while H₂O₂ contributed to anoikis resistance in metastatic melanoma cells. Full article

Tamarindus indica L., a key economic tree species in tropical regions, suffers severely from postharvest decay. From 2023 to 2025, disease fruits exhibiting pericarp softening, pulp browning, and sticky exudates were collected in Yunnan, China. Pathogenicity tests following Koch’s postulates, combined with morphological characterization and phylogenetic analyses of the internal transcribed spacer (ITS), translation elongation factor 1-alpha (TEF 1α), and beta-tubulin (TUB) gene regions, identified the causal pathogen as Botryosphaeria fabicerciana (isolates ZWML-06, ZWML-44, ZWML-17). This is the first report of this postharvest disease on tamarind in Yunnan, filling an etiological gap. Additionally, an endophytic bacterium, designated BV-1, was isolated from asymptomatic pulp tissues. Whole-genome sequencing and phylogenetic analysis identified it as Bacillus velezensis. Strain BV-1 exhibited strong in vitro antagonistic activity against the pathogen, indicating promising biocontrol potential. Functional annotation revealed that BV-1 possesses a complex genetic system with developed transporter systems; its core metabolic network is dominated by nitrogen metabolism and redox processes, suggesting a potential “multi-target” antimicrobial mechanism. This study provides a theoretical basis and novel resources for the green control of postharvest diseases in tamarind. Full article

Hepatocellular carcinoma (HCC) remains a leading cause of cancer-related mortality worldwide, characterized by marked molecular heterogeneity, late-stage diagnosis, and limited therapeutic options. Emerging evidence highlights the interplay between cytoskeletal dynamics, epigenetic regulation, and oncogenic signaling pathways in hepatocarcinogenesis. Histone deacetylase 6 (HDAC6), a key regulator of cytoplasmic protein acetylation, modulates α-tubulin stability, while CTNNB1 (β-catenin) serves as a central effector of the Wnt signaling pathway. However, the existence and functional relevance of a coordinated HDAC6–TUBA1A–CTNNB1 regulatory axis in HCC remain insufficiently explored. We conducted a comprehensive integrative bioinformatic analysis using multiple publicly available datasets and platforms, including TCGA, GEO, GEPIA3, TNMplot, UALCAN, TIMER2.0, STRING, ENCORI, HPA, TargetScan, miRDB, CRISPRdb, GSCALite, and exoRBase. Gene expression, promoter methylation, survival associations, immune infiltration, regulatory RNA interactions, and therapeutic targetability were systematically evaluated. HDAC6 expression was significantly downregulated in HCC tissues, whereas TUBA1A and CTNNB1 were upregulated. Reduced HDAC6 expression was associated with poorer survival outcomes, while TUBA1A and CTNNB1 showed no significant prognostic value. Methylation analysis revealed gene-specific epigenetic alterations, including hypomethylation of CTNNB1 and differential methylation patterns in HDAC6 and TUBA1A. Immune infiltration analysis demonstrated that HDAC6 expression positively correlated with cytotoxic immune cell populations and negatively with immunosuppressive subsets. Regulatory network analyses identified lncRNA–miRNA–mRNA interactions, particularly involving SNHG1. Furthermore, in silico CRISPR targetability and extracellular vesicle (EV) transcript profiling suggested potential translational applicability of this axis. Our findings support a hypothesis of the existence of a dysregulated HDAC6–α-tubulin–β-catenin axis in HCC, linking cytoskeletal remodeling with oncogenic signaling and immune modulation. This axis may indicate a promising candidate for biomarker development and targeted therapeutic strategies, warranting further experimental validation. Full article

Pancreatic cancer (PC) usually results in poor survival with limited treatment options. Reactive oxygen species (ROS) play a key role in the action of HDAC inhibitors. In combination with a ROS generator, it can increase sensitivity to HDAC inhibitors and also overcome resistance to HDAC inhibitors. NQO1-bioactivatable drugs are efficient ROS generators. Therefore, to reduce HDAC inhibitor resistance and enhance its anti-pancreatic cancer activity, in this study, we reported a novel NQO1/HDAC dual-targeting agent, STY12, which exhibited potent anti-pancreatic cancer activity through ROS-mediated DNA damage. STY12 strongly inhibited HDAC1 and HDAC6 activity (IC₅₀ = 29 nM and 10 nM, respectively) and exhibited excellent reduction rates by NQO1 (k_cat/K_m = 5.59 × 10⁶ M⁻¹s⁻¹). STY12 showed good anti-proliferative effects on PC MIA PaCa-2, SW1990, and Capan-2 cells, with IC₅₀ values of 0.23 ± 0.01, 0.25 ± 0.01 and 0.14 ± 0.02 μM, respectively, and lower anti-proliferative effects against normal hTERT-HPNE and BEAS-2B cells. Mechanistic analysis revealed that STY12 suppressed the proliferation of MIA PaCa-2 cells by increasing the acetylation of histone 3 (H3) and α-tubulin, and increasing ROS-mediated DNA damage. Moreover, STY12 arrested the cell cycle at the S phase, inhibited the metastasis of MIA PaCa-2 cells, and promoted their apoptosis. Moreover, compared to SAHA and β-Lap, STY12 exhibited prominent in vivo antitumor activity with negligible toxic effects. Therefore, STY12 can serve as an effective NQO1/HDAC dual-targeting agent for the treatment of PC. Full article